Identification of cellular pathways that regulate Zika virus (ZIKV) replication and pathogenesis is important to our understanding of virus-host interactions, and for the discovery of exploitable cellular drug targets. ZIKV poses an ongoing threat to public health owing to a lack of vaccines and therapeutics, ineffective mosquito control, and the expanding habitats of vectors Aedes aegypti and Aedes albopictus as the result of climate change. Women of child-bearing age are particularly vulnerable, with vertical transmission during pregnancy a significant problem, resulting in devastating consequences to the fetus such as brain anomalies and microcephaly. ZIKV can also be transmitted sexually, with virus observed in the sexual tissues of men and women weeks to months after initial infection. Significant effort is required to understand ZIKV replication and pathogenesis, as well as to treat and prevent infection, including novel means of diagnosis, pharmacotherapy and vaccine development. Cellular factors serve a variety of processes including viral entry, uncoating, fusion, membrane-associated RNA replication, ER-Golgi transport and others. In this proposal a combined chemical- genetic approach will be used to identify key cellular protein kinase pathways that control ZIKV infection. Small molecule and siRNA screens will be performed to identify kinase inhibitors and genes that either positively or negatively regulate ZIKV. Data from the screens will be analyzed to identify cellular pathways that can serve as drug targets and which will deepen our understanding of ZIKV host-pathogen interactions. Preliminary data has identified specific genes and protein kinase targets that have an antiviral function. To compete this focused stage of the project, screens will be conducted to reveal proviral genes and kinases. Selected antiviral and proviral genes will be analyzed using secondary image-based assays for their effects on viral entry, establishment of viral replication centers, and the process of cell-to-cell spread. The resultant data will be prepared for publication along with those for identification of the antiviral genes and kinases. !